Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 46.801
Filtrar
1.
J Vis Exp ; (182)2022 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-35499358

RESUMO

Over the past several decades, biodegradable materials have been extensively explored for biomedical applications such as orthopedic, dental, and craniomaxillofacial implants. To screen biodegradable materials for biomedical applications, it is necessary to evaluate these materials in terms of in vitro cell responses, cytocompatibility, and cytotoxicity. International Organization for Standardization (ISO) standards have been widely utilized in the evaluation of biomaterials. However, most ISO standards were originally established to assess the cytotoxicity of nondegradable materials, thus providing limited value for screening biodegradable materials. This article introduces and discusses three different culture methods, namely, direct culture method, direct exposure culture method, and exposure culture method for evaluating the in vitro cytocompatibility of biodegradable implant materials, including biodegradable polymers, ceramics, metals, and their composites, with different cell types. Research has shown that culture methods influence cell responses to biodegradable materials because their dynamic degradation induces spatiotemporal differences at the interface and in the local environment. Specifically, the direct culture method reveals the responses of cells seeded directly on the implants; the direct exposure culture method elucidates the responses of established host cells coming in contact with the implants; and the exposure culture method evaluates the established host cells that are not in direct contact with the implants but are influenced by the changes in the local environment due to implant degradation. This article provides examples of these three culture methods for studying the in vitro cytocompatibility of biodegradable implant materials and their interactions with bone marrow-derived mesenchymal stem cells (BMSCs). It also describes how to harvest, passage, culture, seed, fix, stain, characterize the cells, and analyze postculture media and materials. The in vitro methods described in this article mimic different scenarios of the in vivo environment, broadening the applicability and relevance of in vitro cytocompatibility testing of different biomaterials for various biomedical applications.


Assuntos
Implantes Absorvíveis , Ortopedia , Terapia Comportamental , Materiais Biocompatíveis , Técnicas de Cultura , Materiais Dentários
2.
Planta ; 255(6): 117, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35513731

RESUMO

MAIN CONCLUSION: The use of beneficial microorganisms improves the performance of in vitro - cultured plants through the improvement of plant nutrition, the biological control of microbial pathogens or the production of phytohormones that promote plant growth and development. Plant in vitro culture techniques are highly useful to obtain significant amounts of true-to-type and disease-free plant materials. One of these techniques is clonal micropropagation which consists on the establishment of shoot tip cultures, shoot multiplication, in vitro rooting and acclimatization to ex vitro conditions. However, in some cases, the existence of recalcitrant genotypes, with a compromised multiplication and rooting ability, or the difficulties to overcome the overgrowth of endophytic contaminations might seriously limit its efficiency. In this sense, the establishment of beneficial interactions between plants and plant growth-promoting microorganisms (PGPMs) under in vitro culture conditions might represent a valuable approach to efficiently solve those restrictions. During the last years, significant evidence reporting the use of beneficial microorganisms to improve the yield of in vitro multiplication or rooting as well as their acclimatization to greenhouse or soil conditions have been provided. Most of these positive effects are strongly linked to the ability of these microorganisms to provide in vitro plants with nutrients such as nitrogen or phosphorous, to produce plant growth regulators, to control the growth of pathogens or to mitigate stress conditions. The culture of A. thaliana under aseptic conditions has provided high-quality knowledge on the root development signaling pathways, involving hormones, triggered in the presence of PGPMs. Overall, the present article offers a brief overview of the use of microorganisms to improve in vitro plant performance during the in vitro micropropagation stages, as well as the main mechanisms of plant growth promotion associated with these microorganisms.


Assuntos
Desenvolvimento Vegetal , Raízes de Plantas , Meios de Cultura , Técnicas de Cultura/métodos , Reguladores de Crescimento de Plantas , Brotos de Planta
3.
Methods Mol Biol ; 2429: 475-484, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35507182

RESUMO

Breast cancer is the most common malignancy worldwide in females, representing 29% of all cancer new cases and 14% of cancer deaths in the world. Amongst the reasons for the high mortality rate is resistance to chemotherapy resulting in therapeutic failure. Various studies have shown that the presence of cancer stem cells (CSCs) in breast tumors is responsible for chemotherapy resistance and tumor recurrence. This CSC population possesses the characteristics of normal stem cells, including their ability to self-renewal and give rise to other epithelial cells. One thing that unique to the CSC population is their ability to escape from chemotherapy drugs; this can make them resistant to therapy and able to repopulate the cancer. Isolation and enrichment of breast CSCs (BCSCs) is required in order to study their characteristics and the behavior that enables them to drive breast tumor development, in order to develop better therapies. This chapter describes a method for the isolation and enrichment of BCSCs from the MCF7 breast cancer cell line, which consists of a heterogeneous breast cancer cell population. This method depends on cancer stem cell behavior, specifically an ability to self-renew and form spheroids in harsh conditions that allow only cancer cells with stem cell characteristics to survive and form spheroids.


Assuntos
Neoplasias da Mama , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Técnicas de Cultura , Feminino , Humanos , Células MCF-7 , Recidiva Local de Neoplasia/patologia , Células-Tronco Neoplásicas/metabolismo , Esferoides Celulares/metabolismo
4.
Int J Mol Sci ; 23(3)2022 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-35163763

RESUMO

The root tissues play important roles in water and nutrient acquisition, environmental adaptation, and plant development. In this study, a diversity panel of 388 wheat accessions was collected to investigate nine root system architecture (RSA) traits at the three-leaf stage under two growing environments: outdoor pot culture (OPC) and indoor pot culture (IPC). Phenotypic analysis revealed that root development was faster under OPC than that under IPC and a significant correlation was observed between the nine RSA traits. The 660K single-nucleotide polymorphism (SNP) chip was used for a genome-wide association study (GWAS). Significant SNPs with a threshold of -log10 (p-value) ≥ 4 were considered. Thus, 36 quantitative trait loci (QTLs), including 13 QTL clusters that were associated with more than one trait, were detected, and 31 QTLs were first identified. The QTL clusters on chromosomes 3D and 5B were associated with four and five RSA traits, respectively. Two candidate genes, TraesCS2A01G516200 and TraesCS7B01G036900, were found to be associated with more than one RSA trait using haplotype analysis, and preferentially expressed in the root tissues. These favourable alleles for RSA traits identified in this study may be useful to optimise the root system in wheat.


Assuntos
Mapeamento Cromossômico/métodos , Estudo de Associação Genômica Ampla/métodos , Locos de Características Quantitativas , Triticum/crescimento & desenvolvimento , Técnicas de Cultura , Desequilíbrio de Ligação , Fenótipo , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/genética , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Polimorfismo de Nucleotídeo Único , Triticum/genética
5.
Mar Drugs ; 20(2)2022 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-35200614

RESUMO

Marine microorganisms have proven to be a source of new natural products with a wide spectrum of biological activities relevant in different industrial sectors. The ever-increasing number of sequenced microbial genomes has highlighted a discrepancy between the number of gene clusters potentially encoding the production of natural products and the actual number of chemically characterized metabolites for a given microorganism. Homologous and heterologous expression of these biosynthetic genes, which are often silent under experimental laboratory culture conditions, may lead to the discovery of new cryptic natural products of medical and biotechnological interest. Several new genetic and cultivation-based strategies have been developed to meet this challenge. The OSMAC approach (one strain-many compounds), based on modification of growth conditions, has proven to be a powerful strategy for the discovery of new cryptic natural products. As a direct extension of this approach, the addition of chemical elicitors or epigenetic modifiers have also been used to activate silent genes. This review looks at the structures and biological activities of new cryptic metabolites from marine-derived microorganisms obtained using the OSMAC approach, the addition of chemical elicitors, and enzymatic inhibitors and epigenetic modifiers. It covers works published up to June 2021.


Assuntos
Organismos Aquáticos/microbiologia , Produtos Biológicos/farmacologia , Descoberta de Drogas/métodos , Animais , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Biotecnologia/métodos , Técnicas de Cultura , Epigênese Genética , Humanos
6.
J Oleo Sci ; 71(1): 119-125, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35013034

RESUMO

Moesziomyces antarcticus is a basidiomycetous yeast that produces mannosylerythritol lipids (MELs), which have potential applications as bio-based functional materials in various oleochemical industries, the cosmetics, toiletry, agriculture, and pharmaceutical industries. To better understand the MEL producer, we characterized the central metabolic pathways of M. antarcticus strain T-34 grown on glucose or olive oil via metabolomics. The relative fatty acid content was higher in the cells cultured in olive oil compared to glucose, while the acetyl-CoA content was lower in cells cultured in olive oil. The levels of the tricarboxylic acid cycle metabolites citrate/isocitrate, α-ketoglutarate, and succinate were lower in olive oil compared to glucose, while fumarate and malate levels exhibited the opposite pattern. Pyruvate was not detected in olive oil compared to glucose culture. The levels of glycerol, as well as trehalose, myo-inositol, threitol/erythritol, and mannitol/sorbitol, were higher in olive oil compared to glucose cultures. The ATP level was lower in olive oil compared to glucose culture, although the assimilation of fatty acids produced by digestion of olive oil should promote large amounts of ATP production. The possibility that ATP regeneration by respiratory chain complex promote oil utilization and MEL production in M. antarcticus T-34 was found based on the results of this metabolomic analysis.


Assuntos
Basidiomycota/metabolismo , Glicolipídeos/biossíntese , Redes e Vias Metabólicas/fisiologia , Metabolômica/métodos , Acetilcoenzima A/metabolismo , Trifosfato de Adenosina/metabolismo , Ciclo do Ácido Cítrico , Meios de Cultura , Técnicas de Cultura , Ácidos Graxos/metabolismo , Fumaratos/metabolismo , Glucose , Glicerol/metabolismo , Malatos/metabolismo , Azeite de Oliva
7.
Sci Rep ; 12(1): 1251, 2022 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-35075262

RESUMO

Staphylococcus aureus is an opportunistic, pathogenic bacteria that causes significant morbidity and mortality. As antibiotic resistance by S. aureus continues to be a serious concern, developing novel drug therapies to combat these infections is vital. Quorum sensing inhibitors (QSI) dampen S. aureus virulence and facilitate clearance by the host immune system by blocking quorum sensing signaling that promotes upregulation of virulence genes controlled by the accessory gene regulator (agr) operon. While QSIs have shown therapeutic promise in mouse models of S. aureus skin infection, their further development has been hampered by the suggestion that agr inhibition promotes biofilm formation. In these studies, we investigated the relationship between agr function and biofilm growth across various S. aureus strains and experimental conditions, including in a mouse model of implant-associated infection. We found that agr deletion was associated with the presence of increased biofilm only under narrow in vitro conditions and, crucially, was not associated with enhanced biofilm development or enhanced morbidity in vivo.


Assuntos
Proteínas de Bactérias/fisiologia , Biofilmes/crescimento & desenvolvimento , Staphylococcus aureus/fisiologia , Transativadores/fisiologia , Animais , Técnicas de Cultura , Feminino , Camundongos Endogâmicos BALB C , Percepção de Quorum
8.
Int J Environ Health Res ; 32(3): 616-627, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32627584

RESUMO

As therapeutic antiviral agents, biological nanoparticles can fight the drug-resistant types of viruses helping the antiviral drug development. In this study, two blue-green algal strains; Oscillatoria sp. and Spirulina platensis were used, mediated by green Ag2O|AgO-NPs and Au-NPs, respectively. For NPs characterization, the UV/Vis spectroscopy were used where their formation and crystallinity were proven with λmax values for silver and gold NPs of 432 and 552 nm, respectively. The transmission electron microscope (TEM) X-ray diffraction showed a spherical-shaped Ag2O|AgO-NPs (size; 14.42 to 48.97) while Au-NPs appeared with octahedral, pentagonal and triangular structures (size; 15.60-77.13 nm). The reducing, capping, and stabilization activities of algal polysaccharides and proteins were indicated via FTIR spectroscopy. Both Ag2O|AgO-NPs and Au-NPs were investigated against Herpes Simplex virus (HSV-1) that has been indicated by its reduction activity of cytopathic effect (CPE). Cytotoxicity was evaluated on Vero cells and measured by MTT assay. Results showed a 90% reduction in CPE of HSV-1 applying Ag2O|AgO-NPs, and Au-NPs at 31.25 µL., with a high reduction rate (49.23%) with Ag2O|AgO-NPs than that of Au-NPs (42.75%). Current results proved the efficiency of green nanotechnology application with both Ag2O|AgO-NPs, and Au-NPs as reducing and inhibitory agents for the HSV-1 replication.


Assuntos
Herpesvirus Humano 1 , Nanopartículas Metálicas , Animais , Antivirais/farmacologia , Chlorocebus aethiops , Técnicas de Cultura , Ouro , Prata , Células Vero
9.
Mycoses ; 65(1): 24-29, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34181777

RESUMO

BACKGROUND: Aspergillus species is the most common agent of invasive pulmonary fungal disease. Culture-based diagnosis considered as gold standard is limited by the fungal load in samples. Detection of Aspergillus by polymerase chain reaction (PCR) has been included as a diagnostic criterion by European Organisation for Research and Treatment of Cancer (EORTC). Most routine laboratories lack facilities for molecular diagnosis. Better yield using high-volume culture (HVC) technique has been reported. Studies have not compared HVC and PCR for detection of Aspergillus species in respiratory samples from patients with suspected invasive pulmonary Aspergillosis (IPA) not on antifungal therapy. OBJECTIVE: This pilot study compared HVC and PCR for the detection of Aspergillus species in respiratory samples from treatment naïve patients. METHODS: Bronchoalveolar lavage (BAL) samples from 30 patients with clinical suspicion of IPA were evaluated. Direct microscopy, culture both conventional (CC) and HVC and qualitative Pan Aspergillus PCR were performed. Latent class model was used for statistical analysis. RESULTS: Sensitivity of HVC (100%) was better compared with CC (60%) and comparable to that of PCR (100%). Specificities of CC, HVC and PCR were 100%, 100% and 25%, respectively. CONCLUSION: High-volume culture is a simple cost-effective technique with a high sensitivity and specificity. It can be easily introduced in routine microbiology laboratories. In centres with the availability of infrastructure for molecular analysis, Aspergillus PCR with other mycological techniques can be used for better diagnosis and management of patients with IPA.


Assuntos
Técnicas de Cultura , Aspergilose Pulmonar Invasiva , Aspergillus/genética , Líquido da Lavagem Broncoalveolar/microbiologia , DNA Fúngico/genética , Humanos , Aspergilose Pulmonar Invasiva/diagnóstico , Projetos Piloto , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
10.
Eur Rev Med Pharmacol Sci ; 25(1 Suppl): 101-107, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34890040

RESUMO

OBJECTIVE: The aim of the study was to show the importance of developing techniques that could exploit the potential of bacteriophages as therapeutics or food supplements. MATERIALS AND METHODS: PubMed database was searched using the following combination of keywords: (bacteriophage) AND (human therapy); (natural bacteriophage) AND (application). RESULTS: The increasing antibiotic resistance of many bacterial strains is making standard antibiotic treatments less effective. Phage therapy provides a non-antibiotic alternative with greater specificity and without harmful effects on the human microbiota. Phages target their specific bacteria, replicate, and then, destroy the host pathogen. Bacteriophages may be administered by several routes, including topical, oral and intravenous. They not only destroy the host pathogen but, in some cases, increase the sensitivity of host bacteria to antibiotics. Various studies have shown that combining phage therapy and antibiotic treatment can be effective against bacterial infections. Clinical trials of phage therapy have shown promising results for various human diseases and conditions. With advances in genetic engineering and molecular techniques, bacteriophages will be able to target a wide range of bacteria. CONCLUSIONS: In the future, phage therapy promises to become an effective therapeutic option for bacterial infections. Since many potentially beneficial bacteriophages can be found in food, supplements containing bacteriophages could be designed to remodel gut microbiota and eliminate pathogenic bacteria. Remodeling of gut microbiota could correct gut dysbiosis. The order of phages known to have these promising activities is Caudovirales, especially the families Siphoviridae and Myoviridae.


Assuntos
Infecções Bacterianas/terapia , Bacteriófagos , Terapia por Fagos/métodos , Infecções Bacterianas/fisiopatologia , Infecções Bacterianas/virologia , Bacteriófagos/isolamento & purificação , Bacteriófagos/fisiologia , Técnicas de Cultura/métodos , Técnicas de Cultura/tendências , Disbiose/fisiopatologia , Disbiose/terapia , Disbiose/virologia , Microbioma Gastrointestinal/fisiologia , Humanos , Terapia por Fagos/tendências
11.
Sci Rep ; 11(1): 24311, 2021 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-34934139

RESUMO

The planarian species Schmidtea mediterranea is a flatworm living in freshwater that is used in the research laboratory as a model to study developmental and regeneration mechanisms, as well as antibacterial mechanisms. However, the cultivable microbial repertoire of the microbes comprising its microbiota remains unknown. Here, we characterized the bacterial constituents of a 10-year-old laboratory culture of planarian species S. mediterranea via culturomics analysis. We isolated 40 cultivable bacterial species, including 1 unidentifiable species. The predominant phylum is Proteobacteria, and the most common genus is Pseudomonas. We discovered that parts of the bacterial flora of the planarian S. mediterranea can be classified as fish pathogens and opportunistic human pathogens.


Assuntos
Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Laboratórios/estatística & dados numéricos , Microbiota , Planárias/microbiologia , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Técnicas de Cultura , RNA Ribossômico 16S/genética , Regeneração , Manejo de Espécimes
12.
PLoS One ; 16(12): e0260645, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34941870

RESUMO

Conventional in vitro culture and manipulation of mouse embryos require a CO2 incubator, which not only increases the cost of performing experiments but also hampers the transport of embryos to the other laboratories. In this study, we established and tested a new CO2 incubator-free embryo culture system and transported embryos using this system. Using an Anaero pouch, which is a CO2 gas-generating agent, to increase the CO2 partial pressure of CZB medium to 4%-5%, 2-cell embryos were cultured to the blastocyst stage in a sealed tube without a CO2 incubator at 37°C. Further, the developmental rate to blastocyst and full-term development after embryo transfer were comparable with those of usual culture method using a CO2 incubator (blastocyst rate: 97% versus 95%, respectively; offspring rate: 30% versus 35%, respectively). Furthermore, using a thermal bottle, embryos were reliably cultured using this system for up to 2 days at room temperature, and live offspring were obtained from embryos transported in this simple and very low-cost manner without reducing the offspring rate (thermal bottle: 26.2% versus CO2 incubator: 34.3%). This study demonstrates that CO2 incubators are not essential for embryo culture and transportation and that this system provides a useful, low-cost alternative for mouse embryo culture and manipulation.


Assuntos
Blastocisto/fisiologia , Dióxido de Carbono/administração & dosagem , Técnicas de Cultura Embrionária/métodos , Transferência Embrionária/métodos , Embrião de Mamíferos/citologia , Animais , Meios de Cultura , Técnicas de Cultura/métodos , Embrião de Mamíferos/fisiologia , Feminino , Fertilização In Vitro , Incubadoras/estatística & dados numéricos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR
13.
Molecules ; 26(22)2021 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-34833914

RESUMO

Linnaea borealis L. (Twinflower)-a dwarf shrub in the Linnaeeae tribe of Caprifoliaceae family-is distributed across the Northern Hemisphere. By means of this study, a reliable protocol for efficient micropropagation of uniform L. borealis L. var. borealis plantlets has been provided for the first time; callus culture was also established. Different initial explants, types of cultures, media systems, and plant growth regulators in Murashige and Skoog (MS) media were tested. Agitated shoot cultures in the liquid media turned out to be the best system for the production of sustainable plant biomass. After stabilization of the callus lines, the highest growth index (c.a. 526%) was gained for callus maintained on MS enriched with picloram. TLC and UHPLC-HESI-HRMS analysis confirmed the presence of phenolic acids and flavonoids, and for the first time, the presence of iridoids and triterpenoid saponins in this species. Multiplication of L. borealis shoot culture provides renewable raw material, allowing for the assessment of the phytochemical profile, and, in the future, for the quantitative analyses and the studies of the biological activity of extracts, fractions, or isolated compounds. This is the first report on in vitro cultures of traditionally used L. borealis rare taxon and its biosynthetic potential.


Assuntos
Caprifoliaceae/química , Caprifoliaceae/crescimento & desenvolvimento , Compostos Fitoquímicos/química , Biomassa , Caprifoliaceae/genética , Conservação dos Recursos Naturais/métodos , Meios de Cultura , Técnicas de Cultura , Flavonoides/química , Genoma de Planta , Horticultura/métodos , Iridoides/química , Saponinas/química , Triterpenos/química
14.
J Reprod Dev ; 67(5): 327-331, 2021 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-34483145

RESUMO

Mammalian ovaries contain a large number of immature follicles. Follicular culture can contribute to the production of fertile oocytes from latent immature follicles, providing a useful tool for exploring the developmental competencies and related factors that oocytes acquire during growth. However, the potential of oocytes produced by follicular culture is limited. Herein, the optimal follicular culture conditions for the addition of polyvinylpyrrolidone to the medium and oxygen concentration were investigated. Polyvinylpyrrolidone with a high molecular weight (≥ 360,000) and a 7% oxygen concentration were found to increase the blastocyst formation rate by more than 20% compared with conventional culture conditions. Although the developmental ability of oocytes produced by follicular culture remained inferior to that of in vivo-derived oocytes, these findings may pave the way for enhanced production of fertile oocytes in vitro and for studying the process of full developmental potency acquisition by oocytes.


Assuntos
Técnicas de Cultura , Oócitos/crescimento & desenvolvimento , Folículo Ovariano/efeitos dos fármacos , Povidona/administração & dosagem , Animais , Feminino , Camundongos
15.
Reprod Biomed Online ; 43(4): 587-597, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34474974

RESUMO

Isolation and characterization of presumptive human adult ovarian stem cells (OSC) has broken the long standing dogma of the absence of postnatal neo-oogenesis. Human adult OSC have been immunosorted by antibodies reacting against the RNA helicase VASA and have been reported to engraft into appropriate stem cell niches to promote neo-oogenesis. Analysis of published research, however, questions some of the findings on isolation, characterization, in-vitro self-renewal and clinical safety of the presumptive human adult OSC. In the present study, human VASApos embryo-fetal primordial germ cells and presumptive adult OSC are shown to share several pluripotency and early germ cell markers not ascertained in the initial characterization of adult OSC. A new hypothesis is made that the restoration of fertility claimed to result from presumptive human adult OSC may be attributed instead to VASApos embryo-fetal primordial germ cell remnants in the adult ovary, or alternatively to earlier VASAneg germ cells generated by in-vitro de-differentiation of the presumptive OSC. The suggested hypotheses have extensive implications for the practice and safety of adult OSC in the development of new treatments aimed at rescuing the ovarian reserve.


Assuntos
Células-Tronco Adultas/enzimologia , RNA Helicases DEAD-box/metabolismo , Células Germinativas/enzimologia , Infertilidade Feminina/terapia , Ovário/citologia , Transplante de Células-Tronco , Animais , Biomarcadores/metabolismo , Separação Celular , Técnicas de Cultura , Embrião de Mamíferos/citologia , Feminino , Humanos
16.
Am J Trop Med Hyg ; 105(6): 1657-1661, 2021 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-34544041

RESUMO

Standard diagnostics for Mycobacterium tuberculosis (MTB) including acid-fast bacilli (AFB) smear and culture, and Xpert™ MTB/RIF real-time Polymerase Chain Reaction (RT-PCR; Xpert) have variable sensitivity and/or long turnaround times. We describe the clinical performance of a laboratory-developed tissue-based MTB PCR compared with AFB culture and Xpert using a composite reference standard (CRS). Over an 8-year period, MTB PCR was performed on pulmonary, pleural, or lymph node specimens for 36 patients. Of these, 11 met criteria for confirmed/probable MTB using CRS. MTB PCR was positive in 100% (11/11), AFB cultures were positive in 73% (8/11), and Xpert in 0% (0/4). MTB PCR was negative in 25 cases of "No MTB" (100% specific). The MTB PCR assay resulted faster than positive AFB culture (mean time 4.3 versus 21.2 days). Tissue-based MTB PCR was associated with increased and rapid detection of MTB, improving clinical sensitivity in strongly suspected MTB cases.


Assuntos
Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Tuberculose dos Linfonodos/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Tuberculose Pleural/diagnóstico , Tuberculose Pulmonar/diagnóstico , Adulto , Idoso , Técnicas de Cultura , Feminino , Humanos , Pulmão/microbiologia , Linfonodos/microbiologia , Masculino , Pessoa de Meia-Idade , Pleura/microbiologia , Padrões de Referência , Estudos Retrospectivos , Sensibilidade e Especificidade , Escarro/microbiologia , Tuberculose/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/fisiopatologia , Tuberculose Pulmonar/fisiopatologia
19.
Int J Mol Sci ; 22(16)2021 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-34445761

RESUMO

Natural products of microbial origin have inspired most of the commercial pharmaceuticals, especially those from Actinobacteria. However, the redundancy of molecules in the discovery process represents a serious issue. The untargeted approach, One Strain Many Compounds (OSMAC), is one of the most promising strategies to induce the expression of silent genes, especially when combined with genome mining and advanced metabolomics analysis. In this work, the whole genome of the marine isolate Rhodococcus sp. I2R was sequenced and analyzed by antiSMASH for the identification of biosynthetic gene clusters. The strain was cultivated in 22 different growth media and the generated extracts were subjected to metabolomic analysis and functional screening. Notably, only a single growth condition induced the production of unique compounds, which were partially purified and structurally characterized by liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS). This strategy led to identifying a bioactive fraction containing >30 new glycolipids holding unusual functional groups. The active fraction showed a potent antiviral effect against enveloped viruses, such as herpes simplex virus and human coronaviruses, and high antiproliferative activity in PC3 prostate cancer cell line. The identified compounds belong to the biosurfactants class, amphiphilic molecules, which play a crucial role in the biotech and biomedical industry.


Assuntos
Antivirais/metabolismo , Glicolipídeos/metabolismo , Rhodococcus/metabolismo , Animais , Antivirais/análise , Chlorocebus aethiops , Técnicas de Cultura , Ensaios de Seleção de Medicamentos Antitumorais , Ésteres/metabolismo , Genoma Bacteriano , Glicolipídeos/química , Humanos , Metaboloma , Testes de Sensibilidade Microbiana , Estrutura Molecular , Células PC-3 , Rhodococcus/química , Rhodococcus/genética , Succinatos/metabolismo , Tensoativos/química , Tensoativos/metabolismo , Células Vero
20.
Eur J Med Res ; 26(1): 85, 2021 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-34344456

RESUMO

BACKGROUND: Streptococcus suis (Ss) is a Gram-positive and anaerobic zoonotic pathogen that is susceptible to all populations and can cause meningitis, septicemia, endocarditis and arthritis in humans. METHODS: In this study, patients with meningitis who were admitted to our hospital with negative blood and cerebrospinal fluid culture were divided into a next-generation sequencing group and a control group. In the next-generation sequencing group, we used the next-generation sequencing method to detect pathogenic bacteria in the patients' cerebrospinal fluid. In the control group, we used blood and cerebrospinal fluid bacterial culture method to detect pathogenic bacteria in the patients' cerebrospinal fluid. The detection rates of pathogenic bacteria in the cerebrospinal fluid of the two groups were compared and analyzed. RESULTS: A total of 18 patients were included in this study, including 8 patients in the next-generation sequencing group and 10 patients in the control group. The mean age (P = 0.613) and mean disease duration (P = 0.294) were similar in both groups. Patients in the next-generation sequencing group had a leukocyte count of 13.13 ± 4.79 × 109, a neutrophil percentage of 83.39 ± 10.36%, and a C-reactive protein level of 134.95 ± 107.69 mg/L. Patients in the control group had a temperature of 38.32 ± 1.07, a leukocyte count of 8.00 ± 2.99 × 109, and a neutrophil percentage of 74.61 ± 8.89%, and C-reactive protein level was 4.75 ± 6.8 mg/L. The statistical results showed that the leukocytes (P = 0.013) and C-reactive protein levels (P = 0.001) were significantly higher in the patients of the next-generation sequencing group than in the control group. No statistically significant differences were seen in body temperature and neutrophil percentage between the two groups (P > 0.05). The incidence of intracranial pressure and meningeal irritation signs were similar in the two groups (P > 0.05). The detection rate of Streptococcus suis in the cerebrospinal fluid of patients in the next-generation sequencing group was 100%, and the detection rate of Streptococcus suis in the cerebrospinal fluid of the control group was 0%. CONCLUSION: The detection rate of Streptococcus suis infection in cerebrospinal fluid by next-generation sequencing was significantly higher than that by blood and cerebrospinal fluid bacterial culture. Therefore, the diagnosis of porcine streptococcal meningitis by next-generation sequencing method is worthy of clinical promotion and application.


Assuntos
Sangue/microbiologia , Líquido Cefalorraquidiano/microbiologia , Técnicas de Cultura/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Meningites Bacterianas/diagnóstico , Infecções Estreptocócicas/diagnóstico , Streptococcus suis/isolamento & purificação , Animais , Estudos de Casos e Controles , Líquido Cefalorraquidiano/metabolismo , Feminino , Seguimentos , Humanos , Masculino , Meningites Bacterianas/sangue , Meningites Bacterianas/líquido cefalorraquidiano , Meningites Bacterianas/microbiologia , Pessoa de Meia-Idade , Prognóstico , Infecções Estreptocócicas/sangue , Infecções Estreptocócicas/líquido cefalorraquidiano , Infecções Estreptocócicas/microbiologia , Streptococcus suis/genética , Suínos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...